DBCO-PEG4-Butyraldehyde

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Cod Uracil-DNA Glycosylase (Cod UNG) from Atlantic Cod is the only commercially available UNG enzyme that is completely and irreversibly inactivated by moderate heat treatment. The enzyme is produced in a recombinant E. coli (ung-) strain that contains a modified Cod UNG gene.

Key Features:

• Heat-labile – Completely and irreversibly inactivated at 55°C
• Contamination control – ideal in applications below
• Use of Cod UNG makes contamination control possible in RT-PCR
• Does not degrade PCR product post-PCR. This makes downstream use of the PCR product possible
• High purity enzyme, tested free of contaminating nucleases
• Detergent free
• Post-PCR Analysis – Enables post-PCR analysis

Applications

Ideal for contamination control in 

• PCR carry-over prevention
• RT-PCR
• RT-qPCR
• qPCR
• kPCR 
• RT-LAMP

Heat-labile Uracil-DNA Glycosylase

There are several commercially available Uracil-DNA glycosylases on the market today. Most of them are of bacterial origin and work well if you have no intention to further analyze the PCR products post-PCR. However, if you want to store your PCR products for downstream analysis such as cloning and sequencing, the reactivation of UNG and subsequent degradation of your PCR products are a problem with most of the commercially available UNGs. Cod UNG from ArcticZymes is completely and irreversibly inactivated by heat thus ensuring that sample integrity is maintained long-term regardless of storage conditions.

This is illustrated in figure 1, below

Figures

Properties

Recommended Protocols

1. Contamination control in PCR, qPCR and one-step RT-qPCR

Cod UNG works in all commercially available master mixes. 

Be sure that you have used dUTP containing dNTP mixes in your previous PCR experiments.

• Add 0.2 U Cod UNG directly to your 20 µl PCR reaction.
• pre-incubate for 5 min at room temperature.
• For RT-qPCR, reverse transcribe your RNA at 50-55°C.
• Run your PCR.
• Store your PCR product at -20°C or 4°C degrees.

2. Contamination control in RT-LAMP

Cod UNG is ideal for contamination control in RT-LAMP.  
One unit of Cod UNG per 30 μl reaction is sufficient for removing even high concentrations of carry-over contamination.

• Ensure that you use dNTP mixes containing dUTP in your experiments.
• Check that the RT-LAMP reaction is compatible with dUTP by running side-by-side reactions containing different ratios of dUTP to dUTP (100% dUTP, 90% dUTP, 80% dUTP and 0% dUTP).
• Add 1 U Cod UNG directly to your 30 µl RT-LAMP reaction.
• Prepare the reaction mix on ice.
• Analyze your RNA at 65°C, no preincubation is necessary.

Please refer to Protocol for Carry-over Contamination Control

Ordering Info

Cod Uracil-DNA Glycosylase (Cod UNG) from Atlantic Cod is the only commercially available UNG enzyme that is completely and irreversibly inactivated by moderate heat treatment. The enzyme is produced in a recombinant E. coli (ung-) strain that contains a modified Cod UNG gene.

DBCO-PEG24-TFP ester is a heterobifunctional PEG linker with an azide-reactive DBCO and an amine-reactive TFP ester that is less susceptible to hydrolysis than an NHS ester. The PEG linker improves the water solubility of the compound. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

DBCO-PEG24-TFP ester is a heterobifunctional PEG linker with an azide-reactive DBCO and an amine-reactive TFP ester that is less susceptible to hydrolysis than an NHS ester. The PEG linker improves the water solubility of the compound. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.